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Assay Kit Recommendations

Determine the best kit for your application based on sample type, target, and preferred detection method. Cayman’s
Kit Recommendation Guide provides an easy-to-use reference tool to compare Cayman’s assay kits and find the right
one to help make your research possible.

Cell Biology

Assay Kits to Assess Cell Health

Cayman offers a group of related assays that can be used to assess proliferative activity, cell viability,
metabolic activity, cell cycle phase, drug toxicity, and apoptosis. Together, the information derived from
these
assays can indicate whether a cell population that has been exposed to an experimental stimulus is healthy
or
dying, actively dividing or in stasis, or has committed to an apoptotic pathway.

Kit Recommendations for Cyclic AMP and Cyclic GMP Measurement

Cyclic AMP (cAMP) and cyclic GMP (cGMP) are crucial second messengers involved in numerous signal
transduction
pathways. They are linked to various G protein-coupled receptors, cyclic nucleotide-gated channels, or
certain
protein kinases and promote the phosphorylation or dephosphorylation of various downstream enzymes or
cascade of
enzymes, activating or inhibiting the targeted activity. The concentration of cAMP or cGMP in a cell can
serve
as a
marker for the functionality of certain receptors or kinase activity. Cayman offers a panel of cyclic
nucleotide
kits tailored to provide highly sensitive results for accurate determination of cAMP/cGMP levels.

Immunology

Kit Recommendations for Innate Immune Responses: Neutrophil Biology

The first line of defense in the innate immune response to many pathogenic organisms involves the actions of
short-lived neutrophils,
inflammatory cells that are highly adept at ingesting and destroying bacteria. They produce antimicrobial
proteins
and peptides including myeloperoxidase and neutrophil elastase, which are released along with DNA and
histones
to
form extracellular traps, web-like structures that ensnare and kill extracellular pathogens. Cayman’s assay
kits
provide a convenient means to isolate neutrophils and measure the production of antimicrobial products.

Peptidylarginine Deiminases (Protein Arginine Deiminases; PADs) are guanidino-modifying enzymes that
catalyze
the
conversion of specific arginine residues to citrulline. Multiple PAD substrates have been identified,
including
histones, fibrinogen, vimentin, and others. PAD4 itself can undergo autocitrullination at several sites,
which
inhibits its enzymatic activity and may play an important role in regulating citrullination in cells. PAD
activity
is increased in rheumatoid arthritis, producing an abundance of citrulline-containing proteins that can be
recognized by autoantibodies that cause the immune system to attack its own tissues. PADs have also been
implicated
in several other diseases including multiple sclerosis, Alzheimer’s disease, glaucoma, and cancer.

Inflammation

Kit Recommendations for Cyclooxygenase Activity & Inhibition

Cayman Chemical offers several options for measuring cyclooxygenase (COX) activity. A variety of assay kits
are
available to either directly measure prostaglandin F2α (PGF2α) produced in the COX
reaction or to monitor the
peroxidase component of the reaction. Peroxidase activity can be measured fluorometrically or
colorimetrically.
Both
purified human and ovine COX-1 and COX-2 enzymes are available.

Kit Recommendations for Prostaglandin D2

Prostaglandin D2 (PGD2) is very unstable in the presence of serum proteins.
11β-PGF2α, the primary plasma metabolite of PGD2, has historically been measured as a
urinary
marker of PGD2. However, recent studies suggest that measurement of the predominant urinary
metabolite,
tetranor-PGDM, may provide a more accurate index of systemic PGD2 formation.

Sample types not listed may still be assayed but have not been validated by Cayman Scientists

Kit Recommendations for Prostaglandin E2

Prostaglandin E2 (PGE2) is rapidly metabolized in plasma with a half-life of less than
thirty seconds. It is quickly converted in vivo to the inactive metabolite 13,14-dihydro-15-keto
PGE2
and other downstream metabolites.

NOTE: A minimum of a 1:10 dilution is recommended when assaying urine samples

Sample types not listed may still be assayed but have not been validated by Cayman Scientists

Kit Recommendations for Thromboxane B2

A portion of thromboxane B2 (TXB2) measured in urine is of renal origin and does not
represent
systemic TXB2
formation, thus selecting an appropriate assay for urinary TXB2 is dependent on the application.
The
TX
metabolite
11-dehydro TXB2 is better suited for measurement of systemically produced TXB2 in
urine
samples.

Metabolism

Kit Recommendations for Bioenergetics

Cayman offers a group of related assays that can be easily used to study cellular bioenergetics. Designed as
a
set of tools to evaluate mitochondrial toxicity, the kits can function together or as stand-alone assays.
Assays
are available to screen potential inhibitors of the electron transport chain, assess mitochondrial function
in
cells by detecting the rate of oxygen consumption, determine citrate synthase activity, and measure
mitochondrial membrane potential.

Normal cellular function depends in part on the energy derived from glucose, which is involved in the
tightly
controlled metabolic processes of glycolysis, the pentose phosphate pathway, glycogenesis, glycogenolysis,
and
gluconeogenesis. Cellular energy is also stored as lipids, which can be metabolized to fatty acids for use
as a
fuel source, and proteins, which are either catabolized into amino acids for polymerization into new
proteins or
are oxidized to urea and CO2 as an energy source. Energy released by the oxidation of nutrients
undergoes oxidative phosphosphorylation in the mitochondria to generate ATP. Failure to maintain energy
homeostasis leads to various metabolic disorders, the most prominent of which is diabetes mellitus, a
disease
characterized by persistent hyperglycemia. Cayman carries a variety of assay kits to quantify key components
of
these important metabolic pathways.

Oxidative Injury

Kit Recommendations for Antioxidant Detection/Activity

Reactive oxygen species (ROS) are produced during normal cellular metabolism and can result in oxidative
stress.
Cells have both enzymatic and non-enzymatic mechanisms to reduce ROS. Cayman’s assay kits evaluate many of
the
primary antioxidants.

Redox signaling and oxidative stress are important for many physiological and pathological conditions. Small
molecules such as reactive oxygen, nitrogen, and sulfur species work collectively to regulate redox
signaling
via PTM of cysteine residues in the active and allosteric sites of proteins. To better understand the
roles
of these different PTMs, Cayman has developed kits that can be used to specifically detect individual thiol
modifications in living cells.